SPECTROSCOPIC CHARACTERIZATION OF ALBUMIN AND MYOGLOBIN ENTRAPPED IN BULK SOL-GEL GLASSES

被引:112
作者
EDMISTON, PL [1 ]
WAMBOLT, CL [1 ]
SMITH, MK [1 ]
SAAVEDRA, SS [1 ]
机构
[1] UNIV ARIZONA,DEPT CHEM,TUCSON,AZ 85721
关键词
D O I
10.1006/jcis.1994.1119
中图分类号
O64 [物理化学(理论化学)、化学物理学];
学科分类号
070304 ; 081704 ;
摘要
The immobilization of proteins by entrapment in optically clear, porous glasses prepared by sol-gel techniques appears to be a promising approach to optical biosensor development. However, little is known about the physical environment of the immobilized protein or the mechanism(s) of entrapment. In this study, absorbance and fluorescence spectroscopies have been used to characterize the properties of two model proteins, bovine serum albumin (BSA) and horse heart myoglobin (Mb), entrapped in wet sol-gel glass bulks. The fluorescence behavior of dissolved and entrapped BSA in the presence of acid, a chemical denaturant, and a collisional quencher was examined. The results show that a large fraction of the BSA added to the sol is entrapped within the gelled glass in a native conformation. However, the reversible conformational transitions that BSA undergoes in solution are sterically restricted in the gel. In contrast, the native properties of Mb are largely lost upon entrapment, as judged by the changes in the visible absorbance spectra of dissolved and entrapped Mb in acidic solutions. Fluorescence studies of dissolved and entrapped apomyoglobin support this conclusion. (C) 1994 Academic Press, Inc.
引用
收藏
页码:395 / 406
页数:12
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